Silencing of ANKRD12 circRNA induces molecular and functional changes associated with invasive phenotypes.
Biomarkers, Tumor
/ genetics
Breast
/ cytology
Breast Neoplasms
/ pathology
Cell Movement
Cyclin D1
/ metabolism
Exons
/ genetics
G1 Phase Cell Cycle Checkpoints
Gene Expression Regulation, Neoplastic
Gene Silencing
Humans
Lung
/ cytology
Lung Neoplasms
/ pathology
MCF-7 Cells
Neoplasm Invasiveness
/ genetics
Nuclear Proteins
/ genetics
Phenotype
RNA, Circular
/ genetics
RNA, Small Interfering
/ genetics
Transfection
Breast cancer
Cancer invasion
Circular RNA
OCR
OXPHOS
RNA-seq
siRNA
Journal
BMC cancer
ISSN: 1471-2407
Titre abrégé: BMC Cancer
Pays: England
ID NLM: 100967800
Informations de publication
Date de publication:
11 Jun 2019
11 Jun 2019
Historique:
received:
25
07
2018
accepted:
23
04
2019
entrez:
13
6
2019
pubmed:
13
6
2019
medline:
18
12
2019
Statut:
epublish
Résumé
Circular RNAs (circRNAs) that form through non-canonical backsplicing events of pre-mRNA transcripts are evolutionarily conserved and abundantly expressed across species. However, the functional relevance of circRNAs remains a topic of debate. We identified one of the highly expressed circRNA (circANKRD12) in cancer cell lines and characterized it validated it by Sanger sequencing, Real-Time PCR. siRNA mediated silencing of the circular junction of circANKRD12 was followed by RNA Seq analysis of circANKRD12 silenced cells and control cells to identify the differentially regulated genes. A series of cell biology and molecular biology techniques (MTS assay, Migration analysis, 3D organotypic models, Real-Time PCR, Cell cycle analysis, Western blot analysis, and Seahorse Oxygen Consumption Rate analysis) were performed to elucidate the function, and underlying mechanisms involved in circANKRD12 silenced breast and ovarian cancer cells. In this study, we identified and characterized a circular RNA derived from Exon 2 and Exon 8 of the ANKRD12 gene, termed here as circANKRD12. We show that this circRNA is abundantly expressed in breast and ovarian cancers. The circANKRD12 is RNase R resistant and predominantly localized in the cytoplasm in contrast to its source mRNA. We confirmed the expression of this circRNA across a variety of cancer cell lines and provided evidence for its functional relevance through downstream regulation of several tumor invasion genes. Silencing of circANKRD12 induces a strong phenotypic change by significantly regulating cell cycle, increasing invasion and migration and altering the metabolism in cancer cells. These results reveal the functional significance of circANKRD12 and provide evidence of a regulatory role for this circRNA in cancer progression. Our study demonstrates the functional relevance of circANKRD12 in various cancer cell types and, based on its expression pattern, has the potential to become a new clinical biomarker.
Sections du résumé
BACKGROUND
BACKGROUND
Circular RNAs (circRNAs) that form through non-canonical backsplicing events of pre-mRNA transcripts are evolutionarily conserved and abundantly expressed across species. However, the functional relevance of circRNAs remains a topic of debate.
METHODS
METHODS
We identified one of the highly expressed circRNA (circANKRD12) in cancer cell lines and characterized it validated it by Sanger sequencing, Real-Time PCR. siRNA mediated silencing of the circular junction of circANKRD12 was followed by RNA Seq analysis of circANKRD12 silenced cells and control cells to identify the differentially regulated genes. A series of cell biology and molecular biology techniques (MTS assay, Migration analysis, 3D organotypic models, Real-Time PCR, Cell cycle analysis, Western blot analysis, and Seahorse Oxygen Consumption Rate analysis) were performed to elucidate the function, and underlying mechanisms involved in circANKRD12 silenced breast and ovarian cancer cells.
RESULTS
RESULTS
In this study, we identified and characterized a circular RNA derived from Exon 2 and Exon 8 of the ANKRD12 gene, termed here as circANKRD12. We show that this circRNA is abundantly expressed in breast and ovarian cancers. The circANKRD12 is RNase R resistant and predominantly localized in the cytoplasm in contrast to its source mRNA. We confirmed the expression of this circRNA across a variety of cancer cell lines and provided evidence for its functional relevance through downstream regulation of several tumor invasion genes. Silencing of circANKRD12 induces a strong phenotypic change by significantly regulating cell cycle, increasing invasion and migration and altering the metabolism in cancer cells. These results reveal the functional significance of circANKRD12 and provide evidence of a regulatory role for this circRNA in cancer progression.
CONCLUSIONS
CONCLUSIONS
Our study demonstrates the functional relevance of circANKRD12 in various cancer cell types and, based on its expression pattern, has the potential to become a new clinical biomarker.
Identifiants
pubmed: 31185953
doi: 10.1186/s12885-019-5723-0
pii: 10.1186/s12885-019-5723-0
pmc: PMC6558796
doi:
Substances chimiques
ANKRD12 protein, human
0
Biomarkers, Tumor
0
CCND1 protein, human
0
Nuclear Proteins
0
RNA, Circular
0
RNA, Small Interfering
0
Cyclin D1
136601-57-5
Types de publication
Journal Article
Langues
eng
Sous-ensembles de citation
IM
Pagination
565Subventions
Organisme : Qatar Foundation/ Weill Cornell Medicine Qatar
ID : BMRP 1 - Pilot FY17
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